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Optically-Active Nanocrystals for Inner Filter Effect-Based Fluorescence Sensing: Achieving Better Spectral Overlap
摘要: Among various photophysical processes for fluorescence modulation, inner filter effect (IFE) is well-known for its simplicity and no requirement of the distances between the fluorophores and the corresponding modulators. Theoretically, the key to maximize the sensitivity of IFE-based fluorescent assays is to enlarge the overlap between the absorption of the absorber and the excitation/emission of the fluorophores. However, it is difficult to achieve perfect spectral overlap for IFE in conventional organic fluorophores with constant excitation/emission. The emergence of various optically-active nanocrystals greatly revolutionize the IFE-based fluorescent sensing. Therefore in this critical review, we first made an introduction to IFE from the viewpoint of its photophysical process. Particularly, the similarities and differences of IFE and FRET for better understanding of IFE are discussed, and the general rules for verification of IFE are proposed. The use of various optically-active nanocrystals for enhanced IFE-based sensing is especially concerned in this review.
关键词: fluorescence resonance energy transfer,Inner filter effect,spectral overlap,fluorescent sensors,optically-active nanocrystals,tunable spectra
更新于2025-09-10 09:29:36
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A 1,8 Naphthalimide anchor Rhodamine B Based FRET Probe for Ratiometric Detection of Cr3+ion in Living Cells
摘要: A 2-(1,3-dioxo-1H-benzo[de]isoquinolin-2(3H)-yl)acetaldehydeanchored rhodamine B based probe, RDNAP detects Cr3+ ion by fluorescence resonance energy transfer (FRET) process in aqueous buffered acetonitrile media (7:3, v/v). Conversely, conjugation of 2-(1,3-dioxo-6-(piperidin-1-yl)-1H-benzo[de]isoquinolin-2(3H)-yl)acetaldehyde with rhodamine B provides another probe, RDNAP-PY that undergoes Cr3+assisted ratiometric fluorescence and colorimetric change in the same media. RDNAP-PY provides higher FRET efficiency and detects as low as 1.81×10?6 M Cr3+with an association constant, 15.9 × 104 M-1. Other common ions do not interfere. RDNAP-PY efficiently images intracellular Cr3+ in live Hep3B, MCF-7, HeLa, SiHa and HEK 293T cells under fluorescence microscope in a ratiometric and time dependent manner. 1H NMR titration and DFT studies strongly support experimental findings.
关键词: 1,8 Naphthalimide,Live cell imaging,Ratiometric probe,fluorescence resonance energy transfer,DFT calculation
更新于2025-09-09 09:28:46
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Responsive Upconversion Nanoprobe for Background‐Free Hypochlorous Acid Detection and Bioimaging
摘要: Responsive nanoprobes play an important role in bioassay and bioimaging, early diagnosis of diseases and treatment monitoring. Herein, a upconversional nanoparticle (UCNP)-based nanoprobe, Ru@UCNPs, for specific sensing and imaging of hypochlorous acid (HOCl) is reported. This Ru@UCNP nanoprobe consists of two functional components, i.e., NaYF4:Yb, Tm UCNPs that can convert near infrared light-to-visible light as the energy donor, and a HOCl-responsive ruthenium(II) complex [Ru(bpy)2(DNCH-bpy)](PF6)2 (Ru-DNPH) as the energy acceptor and also the upconversion luminescence (UCL) quencher. Within this luminescence resonance energy transfer nanoprobe system, the UCL OFF–ON emission is triggered specifically by HOCl. This triggering reaction enables the detection of HOCl in aqueous solution and biological systems. As an example of applications, the Ru@UCNPs nanoprobe is loaded onto test papers for semiquantitative HOCl detection without any interference from the background fluorescence. The application of Ru@UCNPs for background-free detection and visualization of HOCl in cells and mice is successfully demonstrated. This research has thus shown that Ru@UCNPs is a selective HOCl-responsive nanoprobe, providing a new way to detect HOCl and a new strategy to develop novel nanoprobes for in situ detection of various biomarkers in cells and early diagnosis of animal diseases.
关键词: imaging,nanoprobes,paper-based test strips,bioassay and bioimaging,luminescence resonance energy transfer,selective HOCl detection and imaging
更新于2025-09-04 15:30:14
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Microscopy of the Heart || Optogenetic Tools in the Microscopy of Cardiac Excitation-Contraction Coupling
摘要: Microscopy became a scienti?c investigation method in the seventeenth century with the application of the ?rst build microscopes on biological samples [1, 2]. Soon it became a popular method to stain samples in order to visualise particular (cellular and subcellular) structures [3]. These stains, either based on absorption or ?uorescence, have limitations in respect to their speci?city and are often toxic to cells, which limits investigations to short intervals or even dead samples. In 1987 the idea came up to use a ?uorescent protein that was discovered 25 years before [4], in particular a green ?uorescent protein (GFP) form the medusa Aequorea victoria to label cells and cellular structures [5]. With the sequencing and cloning of GFP, a so-called ‘green revolution’ started, which led to regular usage of ?uorescent proteins as markers or sensors (for details see below) in the majority of cellular research in physiology, microbiology, pharmacology, molecular biology, anatomy, cell biology, biophysics and many other biomedical ?elds. Although the expression of the ?uorescent proteins and their optical investigation can already be regarded as optogenetic tools, this term was only applied when the optical properties of proteins were used to manipulate cells. The best-known example of such a protein is the channelrhodopsin, a light-gated ion channel [6, 7]. When this ion channel is expressed in a membrane and illuminated with light of the appropriate wavelength, the channel will be activated and opened, which results in passive transportation of ions across the membrane and a change of the membrane potential. However, within this chapter we consider both aspects, the observation and the manipulation as optogenetic tools. To use the optogenetic tool, the genes of these proteins need to be transferred into the cells to allow the expression of the protein. For an overview of gene delivery into target cells, see [8].
关键词: channelrhodopsins,cardiac excitation-contraction coupling,F?rster Resonance Energy Transfer,genetically encoded biosensors,Optogenetic tools,microscopy
更新于2025-09-04 15:30:14
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Resonance Energy Transfer in Arbitrary Media: Beyond the Point Dipole Approximation
摘要: In this work, we present a comprehensive theoretical and computational study of donor/acceptor resonance energy transfer (RET) beyond the dipole approximation, in arbitrary inhomogeneous and dispersive media. The theoretical method extends Fo?rster theory for RET between particles (molecules or nanoparticles) to the case where higher multipole transitions in the donor and/or acceptor play a significant role in the energy transfer process. In our new formulation, the energy transfer matrix element is determined by a fully quantum electrodynamic expression, but its evaluation requires only classical electrodynamics calculations. By means of a time domain electrodynamical approach (TED), the matrix element evaluation involves the electric and magnetic fields generated by the donor and evaluated at the position of the acceptor, including fields associated with transition electric dipoles, electric quadrupoles, and magnetic dipoles in the donor, and the acceptor response to the electric and magnetic fields and to the electric field gradient. As an illustration of the benefits of the new formalism, we tested our method with a 512 atom lead sulfide (PbS) quantum dot as the donor/acceptor in vacuum, and with spherical nanoparticles (toy model) possessing designed transition multipoles. This includes an analysis of the effects of interferences between multipoles in the energy transfer rate. The results show important deviations from the conventional Fo?rster dipole theory that are important even in vacuum but that can be amplified by interaction with a plasmonic nanoparticle.
关键词: multipole transitions,resonance energy transfer,plasmonic nanoparticle,quantum electrodynamics,dipole approximation,RET
更新于2025-09-04 15:30:14